CN101701189A - Method for culturing microbial strain for degrading industrial waste water - Google Patents
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- CN101701189A CN101701189A CN200910223757A CN200910223757A CN101701189A CN 101701189 A CN101701189 A CN 101701189A CN 200910223757 A CN200910223757 A CN 200910223757A CN 200910223757 A CN200910223757 A CN 200910223757A CN 101701189 A CN101701189 A CN 101701189A
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Abstract
The invention provides a method for culturing a microbial strain for degrading industrial waste water, which belongs to the field of biotechnology. The method can solve the problems that the existing strain for degrading the industrial waste water has single degradation role and can not degrade compound pollutants. The method for culturing the microbial strain for degrading the industrial waste water comprises the following steps: preparing liquid culture media containing different concentration gradients of target compounds; obtaining a plasmid-containing strain which takes an aniline compound or a nitrobenzene compound as a sole carbon and nitrogen source and takes phenol as a sole carbon source respectively; carrying out acclimatization, culture and identification of plasmid and degradation capability; and purifying for obtaining the plasmid-containing microbial strain for degrading the industrial waste water. The culture method has the advantages of simple process and low cost of the used raw materials, and the cultured strain has the capability of degrading the compound pollutants and has higher efficiency for degrading aniline, phenol and nitrobenzene compounds.
Description
Technical field
The present invention relates to a kind of cultural method of microorganism strains, relate in particular to a kind of cultural method of microbial strain for degrading industrial waste water; Belong to biological technical field.
Background technology
Industrial wastewater pollution be administer in the current ecological protection one extremely important and have challenging work.Poor from organic waste water complicated component, biodegradability in the production processes such as printing and dyeing, pharmacy, agricultural chemicals, process hides, petrochemical complex and foodstuffs industry, serious pollution of water environment, even will weaken the long-term fecundity of hydrobiont etc.The purification treatment technology of existing trade effluent can reduce biological process, chemical method, physical-chemical coupled method and materialization-biological coupling method etc.Wherein biological process has good economy performance, advantages such as non-secondary pollution.
Environmental pollution is serious in the existing trade effluent mainly contains phenol, aniline and nitrobenzene compounds etc.Wherein phenol is the principal pollutant in the trade effluents such as papermaking, coking, oil refining, plastics, weaving.Nitrobenzene compounds is widely used in the production of agricultural chemicals, dyestuff, explosive, rubber and other Chemicals.Nitrobenzene class pollutant in the environment mainly comprises compounds such as oil of mirbane, nitro-chlorobenzene, nitrotoluene, nitrophenols, N-methyl-p-nitroaniline.To utilize microbiological deterioration phenol, aniline and nitrobenzene compounds be a kind of both economical and can not produce the method for secondary pollution, in recent years, from the environment of industrial wastewater pollution, be separated to many can degradation of phenol, the microorganism strains of aniline and nitrobenzene compounds.People such as Sheng Lianxi are at Chinese Journal of Applied Ecology, in July, 2007, the 18th volume, the 7th interim p-nitrophenyl compounds microbiological deterioration progress, and from the acclimation and screening of nitrobenzene compounds degradation bacteria, degradation pathway, degraded encourage, are total to metabolism, chemotaxis and molecular genetics angle, have set forth the biodegradable nearest progress of nitrobenzene compounds.People such as Wen Hongyu are in Xuzhou Normal University's journal (natural science edition) in December, 2003, the 21st volume, the 4th interim separation and the physiology characteristic research of having narrated the degradation of phenol bacterium, though it is by strain culturing, domestication and screen and isolate the bacterium that two strains can degradation of phenol from trade effluent, but this bacterial strain only Pyrogentisinic Acid has Degradation, and the combined pollutant that has aniline and nitrobenzene compounds in the trade effluent is not simultaneously but had degradation capability.
Summary of the invention
The present invention is directed to the defective that prior art exists, a kind of cultural method that combined pollutant is had the microbial strain for degrading industrial waste water of degradation capability is provided.
The objective of the invention is to realize by following technical proposal: a kind of cultural method of microbial strain for degrading industrial waste water, this method may further comprise the steps:
A, aniline, phenol and nitrobenzene compounds in the trade effluent, preparation contains the liquid nutrient medium of the target compound of different concns gradient respectively, wherein aniline and nitrobenzene compounds are unique carbon source and nitrogenous source with it, phenol adds nitrogenous source in addition, also add needed other chemical matrix composition of microorganism strains growth in the described liquid nutrient medium, the partially liq substratum is changed into the selectivity solid medium;
B, the active sludge in the trade effluent joined carry out enrichment in the beef extract-peptone liquid nutrient medium, the liquid nutrient medium that is inoculated into the target compound that contains the different concns gradient is then tamed cultivation, culture temperature is 15-45 ℃, incubation time 6-25 days, domestication is cultivated 2-12 time repeatedly, the bacterial strain that domestication is cultivated carries out the plate streaking separation with the selectivity solid medium, and carry out plasmid extraction, obtaining is sole carbon source and nitrogenous source with aniline or nitrobenzene compound respectively, with phenol be sole carbon source contain the plasmid bacterial strain;
C, the pseudomonas strain is inoculated into carries out enrichment in the beef extract-peptone liquid nutrient medium, be made into competent cell after the enrichment, the three kinds of plasmid bacterial strains and the described competent cell that obtain among the step B are resuspended in the LB liquid nutrient medium, are to hatch 10-45 hour under 10-40 ℃ the condition in temperature; The liquid nutrient medium that joins three kinds of target compounds that contain the different concns gradient is again tamed cultivation, the domestication culture temperature is 15-45 ℃, domestication incubation time 6-25 days, domestication is cultivated 2-12 time repeatedly, domestication is coated the selectivity solid medium after cultivating, be to hatch 10-45 hour under 10-40 ℃ the condition in temperature, carry out plasmid and degradation capability then and identify;
D, with after obtaining bacterial strain among the step C and carrying out purifying, promptly obtain to contain the microbial strain for degrading industrial waste water of plasmid.
The present invention is directed to the kind of pollutent in the waste water, adopt the liquid nutrient medium contain different pollutant kinds that active sludge microorganism is tamed cultivation, that adopts the plate isolation method to obtain can pollutent to be sole carbon source, nitrogenous source and the energy again contains the plasmid bacterium.Add competent host bacterium again, adopt the waste water of different concns to tame, the promotion plasmid shifts, and the selective advantage bacterial classification, finally obtains having the microbial strains that contains plasmid of combined pollutant degradation capability behind the purifying.
Wherein prepare the liquid nutrient medium of the target compound of different concns gradient in the steps A, the concentration of mainly preparing target compound is 0,0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L.The liquid nutrient medium of the target compound of different concns gradient can count microbial strain for degrading industrial waste water with the growth change situation of target compound change in concentration and to the degradation rate of target compound; Needed other chemical matrix composition of microbial strains growth mainly contains elements such as phosphorus, iron, potassium, oxygen, sodium, chlorine, calcium, magnesium.To just change into the selectivity solid medium behind the partially liq substratum interpolation 15-20g agar (calculating) by 1000ml water.
What adopt among the step B is conventional beef extract-peptone liquid nutrient medium, and the prescription of substratum is extractum carnis 3 grams, peptone 10 grams, sodium-chlor 5 grams, agar 15 grams, water 1000ml.Adopt molecular biology method to carry out plasmid extraction, mainly adding the centrifugal back of ethidium bromide by centrifugal collection bacterium, alkaline lysis, phenol extracting or cesium chloride, to obtain with aniline or nitrobenzene compound with ethanol sedimentation be sole carbon source and nitrogenous source, with phenol be sole carbon source contain the plasmid bacterial strain.
Pseudomonas strain (Pseudomonas) among the step C mainly is a pseudomonas putida strain, it is inoculated in the conventional beef extract-peptone liquid nutrient medium carries out enrichment, after the enrichment cell suspension at the 100mmol/L of pH6.0 CaCl
2In, under condition of ice bath, placement is spent the night, and allows it change into competent cell.Be 1 * 10 with bacterial concentration then
7-3 * 10
9Three kinds of cfu/mL contain the plasmid bacterial strain and bacterial concentration is 1 * 10
7-3 * 10
9The competent cell of cfu/mL is resuspended in the LB liquid nutrient medium, wherein the LB liquid nutrient medium is used for cultivating in advance bacterial classification, bacterial classification being increased at double, reach service requirements. the bacterial classification of cultivation generally is can't be at the engineering bacteria of independent survival of external environment and amplification through what transform.The prescription of LB liquid nutrient medium is as follows: Tryptones (Tryptone) 10g/L; Yeast extract (Yeast extract) 5g/L; Sodium-chlor (NaCl) 10g/L; Regulate the pH of this substratum with NaOH, make it reach 7.4.
Adopt the streak plate partition method to carry out purifying among the step D to obtaining bacterial strain.
In the cultural method of above-mentioned microbial strain for degrading industrial waste water, the nitrogenous source in the phenol liquid nutrient medium described in the steps A is that concentration is that 500mg-1500mg/L ammonium nitrate or concentration are a kind of in the 500mg-1500mg/L peptone.Adding concentration in the phenol liquid nutrient medium is that 500mg-1500mg/L ammonium nitrate or concentration are that the 500mg-1500mg/L peptone can improve bacterial strain Pyrogentisinic Acid's degradation efficiency as nitrogenous source, wherein adds concentration and is 500mg-1500mg/L peptone degradation efficiency and be higher than that to add concentration be 500mg-1500mg/L ammonium nitrate.
In the cultural method of above-mentioned microbial strain for degrading industrial waste water, other chemical matrix component is in the liquid nutrient medium described in the steps A: saltpetre: 0.5-1.5g/L; Sodium-chlor: 0.1-1g/L; K
2HPO
43H
2O:0.2g-0.8g/L; MgSO
47H
2O:0.2g-0.8g/L; FeSO
47H
2O 0.005-0.015g/L.
As preferably, the culture temperature that domestication is cultivated among the step B is 20-40 ℃, and incubation time 7-21 days, domestication was cultivated 3-10 time repeatedly.
As preferably, among the step C in the LB liquid nutrient medium incubation temperature be 15-35 ℃, incubation time is 12-36 hour.
As preferably, the culture temperature that domestication is cultivated among the step C is 20-40 ℃, and incubation time 7-21 days, domestication was cultivated 3-10 time repeatedly.
As preferably, to coat selectivity solid medium incubation temperature be 15-35 ℃ to bacterial strain among the step C, and incubation time is 12-36 hour.
In sum, the present invention has the following advantages:
1, the bacterial strain that obtains of cultural method of the present invention has degradation capability to combined pollutant, uses more convenient and scope is wider, is convenient to carry out the large-scale industrialization popularization.
2, cultural method technical process of the present invention is simple, uses the cost of raw material lower, and the bacterial strain of cultivation is higher to the aniline in the trade effluent, phenol and nitrobenzene compounds degradation efficiency.
Description of drawings
Fig. 1 is the microbial strain for degrading industrial waste water growth curve chart that embodiment 1 cultivates.
Fig. 2 is microbial strain for degrading industrial waste water and the pH value corresponding relation figure that embodiment 1 cultivates.
Fig. 3 is that the microbial strain for degrading industrial waste water cultivated of embodiment 1 is to organic tolerability curves figure.
Fig. 4 is the graphic representation of the microbial strain for degrading industrial waste water of embodiment 1 cultivation to organic degraded in the trade effluent.
Fig. 5 is the microbial strain for degrading industrial waste water growth curve chart that embodiment 2 cultivates.
Fig. 6 is microbial strain for degrading industrial waste water and the pH value corresponding relation figure that embodiment 2 cultivates.
Fig. 7 is that the microbial strain for degrading industrial waste water cultivated of embodiment 2 is to organic tolerability curves figure.
Fig. 8 is the graphic representation of the microbial strain for degrading industrial waste water of embodiment 2 cultivations to organic degraded in the trade effluent.
Embodiment
Below by specific embodiment also in conjunction with the accompanying drawings, technical scheme of the present invention is described in further detail; But the present invention is not limited to these embodiment.
At the aniline in the trade effluent, phenol and nitrobenzene compounds, to contain concentration be 0g/L in preparation respectively, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the target compound aniline of different concns gradient, phenol and nitrobenzene compounds liquid nutrient medium, wherein aniline and nitrobenzene compounds are unique carbon source and nitrogenous source with it, and phenol adds nitrogenous source peptone 1000mg/L in addition, also add needed other chemical matrix prescription of microorganism strains growth saltpetre: 0.5g/L in the described liquid nutrient medium; Sodium-chlor: 1g/L; K
2HPO
43H
2O:0.8g/L; MgSO
47H
2O:0.2g/L; FeSO
47H
2O 0.010g/L, and will change into the selectivity solid medium behind the partially liq substratum adding interpolation 15g agar (calculating) by 1000ml water.
Active sludge in the trade effluent joined in the conventional beef extract-peptone liquid nutrient medium carry out enrichment, be inoculated into then that to contain concentration be 0g/L, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the target compound aniline of different concns gradient, phenol and nitrobenzene compounds liquid nutrient medium are tamed cultivation, culture temperature is 20 ℃, incubation time 21 days, and domestication is cultivated 3 times repeatedly, the bacterial strain of domestication cultivation is coated the selectivity solid medium carry out the plate streaking separation, and adopt molecular biology method to carry out plasmid extraction, mainly by centrifugal collection bacterium, alkaline lysis, phenol extracting or cesium chloride add the centrifugal back of ethidium bromide to obtain with aniline or nitrobenzene compound with ethanol sedimentation are sole carbon source and nitrogenous source, with phenol be sole carbon source contain the plasmid bacterial strain.
The pseudomonas strain is inoculated in the conventional beef extract-peptone liquid nutrient medium carries out enrichment, after the enrichment cell suspension at the 100mmol/L of pH6.0 CaCl
2In, under condition of ice bath, placement is spent the night, and allows it change into competent cell.Above-mentioned three kinds of plasmid bacterial strains and the described competent cell that obtains is resuspended in the LB liquid nutrient medium, is to hatch 36 hours under 15 ℃ the condition in temperature; Join again that to contain concentration be 0g/L, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the liquid nutrient medium of three kinds of target compounds of different concns gradient is tamed cultivation, the domestication culture temperature is 20 ℃, domestication incubation time 21 days, and domestication is cultivated 3 times repeatedly, domestication is coated the selectivity solid medium after cultivating, be to hatch 36 hours under 15 ℃ the condition in temperature, select single bacterium colony and adopt conventional method to carry out plasmid to identify, and to its growth change situation with aniline concentration, with the growth change situation of waste strength, degradation capability is identified; To its growth performance,, identify that to the tolerance performance of the organism in the waste water with to organic degradation property in the waste water qualification result as shown in Figure 1, Figure 2, Figure 3 and Figure 4 respectively to the conformability of pH value.
Through after identifying that the standard compliant bacterial strain in back adopts the streak plate partition method to carry out purifying, promptly obtain to contain the microbial strain for degrading industrial waste water of plasmid with above-mentioned.
At the aniline in the trade effluent, phenol and nitrobenzene compounds, to contain concentration be 0g/L in preparation respectively, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the liquid nutrient medium of target compound aniline, phenol and the nitrobenzene compounds of different concns gradient, wherein aniline and nitrobenzene compounds are unique carbon source and nitrogenous source with it, and phenol adds nitrogenous source ammonium nitrate 1000mg/L in addition, also add needed other chemical matrix prescription of microorganism strains growth saltpetre: 1.0g/L in the described liquid nutrient medium; Sodium-chlor: 0.5g/L; K
2HPO
43H
2O:0.5g/L; MgSO
47H
2O:0.5g/L; FeSO
47H
2O 0.005g/L, and will change into the selectivity solid medium behind the partially liq substratum adding interpolation 20g agar (calculating) by 1000ml water.
Active sludge in the trade effluent joined in the conventional beef extract-peptone liquid nutrient medium carry out enrichment, be inoculated into then that to contain concentration be 0g/L, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the target compound aniline of different concns gradient, the liquid nutrient medium of phenol and nitrobenzene compounds is tamed cultivation, culture temperature is 30 ℃, incubation time 14 days, and domestication is cultivated 6 times repeatedly, the bacterial strain of domestication cultivation is coated the selectivity solid medium carry out the plate streaking separation, and adopt molecular biology method to carry out plasmid extraction, mainly by centrifugal collection bacterium, alkaline lysis, phenol extracting or cesium chloride add the centrifugal back of ethidium bromide to obtain with aniline or nitrobenzene compound with ethanol sedimentation are sole carbon source and nitrogenous source, with phenol be sole carbon source contain the plasmid bacterial strain.
The pseudomonas strain is inoculated in the conventional beef extract-peptone liquid nutrient medium carries out enrichment, after the enrichment cell suspension at the 100mmol/L of pH6.0 CaCl
2In, under condition of ice bath, placement is spent the night, and allows it change into competent cell.Above-mentioned three kinds of plasmid bacterial strains and the described competent cell that obtains is resuspended in the LB liquid nutrient medium, is to hatch 24 hours under 25 ℃ the condition in temperature; Join again that to contain concentration be 0g/L, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the liquid nutrient medium of three kinds of target compounds of different concns gradient is tamed cultivation, the domestication culture temperature is 30 ℃, domestication incubation time 14 days, domestication is cultivated 6 times repeatedly, coating the selectivity solid medium after domestication is cultivated, is to hatch 24 hours under 25 ℃ the condition in temperature, selects single bacterium colony and adopts conventional method to carry out plasmid to identify, respectively to its growth performance, to the conformability of pH value, identify qualification result such as Fig. 5 to the tolerance performance of the organism in the waste water with to organic degradation property in the waste water, Fig. 6, Fig. 7 and shown in Figure 8.
Through after identifying that the standard compliant bacterial strain in back adopts the streak plate partition method to carry out purifying, promptly obtain to contain the microbial strain for degrading industrial waste water of plasmid with above-mentioned.
At the aniline in the trade effluent, phenol and nitrobenzene compounds, to contain concentration be 0g/L in preparation respectively, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the liquid nutrient medium of target compound aniline, phenol and the nitrobenzene compounds of different concns gradient, wherein aniline and nitrobenzene compounds are unique carbon source and nitrogenous source with it, and phenol adds nitrogenous source peptone 1000mg/L in addition, also add needed other chemical matrix prescription of microorganism strains growth saltpetre: 1.5g/L in the described liquid nutrient medium; Sodium-chlor: 0.1g/L; K
2HPO
43H
2O:0.2g/L; MgSO
47H
2O:0.8g/L; FeSO
47H
2O 0.015g/L, and will change into the selectivity solid medium behind the partially liq substratum adding interpolation 20g agar (calculating) by 1000ml water.
Active sludge in the trade effluent joined in the conventional beef extract-peptone liquid nutrient medium carry out enrichment, be inoculated into then that to contain concentration be 0g/L, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the target compound aniline of different concns gradient, the liquid nutrient medium of phenol and nitrobenzene compounds is tamed cultivation, culture temperature is 40 ℃, incubation time 7 days, and domestication is cultivated 8 times repeatedly, the bacterial strain of domestication cultivation is coated the selectivity solid medium carry out the plate streaking separation, and adopt molecular biology method to carry out plasmid extraction, mainly by centrifugal collection bacterium, alkaline lysis, phenol extracting or cesium chloride add the centrifugal back of ethidium bromide to obtain with aniline or nitrobenzene compound with ethanol sedimentation are sole carbon source and nitrogenous source, with phenol be sole carbon source contain the plasmid bacterial strain.
The pseudomonas strain is inoculated in the conventional beef extract-peptone liquid nutrient medium carries out enrichment, after the enrichment cell suspension at the 100mmol/L of pH6.0 CaCl
2In, under condition of ice bath, placement is spent the night, and allows it change into competent cell.Above-mentioned three kinds of plasmid bacterial strains and the described competent cell that obtains is resuspended in the LB liquid nutrient medium, is to hatch 12 hours under 35 ℃ the condition in temperature; Join again that to contain concentration be 0g/L, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 2.5g/L, 3.0g/L, 3.5g/L the liquid nutrient medium of three kinds of target compounds of different concns gradient is tamed cultivation, the domestication culture temperature is 40 ℃, domestication incubation time 7 days, and domestication is cultivated 8 times repeatedly, domestication is coated the selectivity solid medium after cultivating, be to hatch 12 hours under 35 ℃ the condition in temperature, select single bacterium colony and adopt conventional method to carry out plasmid to identify, respectively to its growth performance, to the conformability of pH value, identify to the tolerance performance of the organism in the waste water with to organic degradation property in the waste water.
Through after identifying that the standard compliant bacterial strain in back adopts the streak plate partition method to carry out purifying, promptly obtain to contain the microbial strain for degrading industrial waste water of plasmid with above-mentioned.
Specific embodiment described in the present invention only is that the present invention's spirit is illustrated.The technician of the technical field of the invention can make various modifications or replenishes or adopt similar mode to substitute described specific embodiment, but can't depart from spirit of the present invention or surmount the defined scope of appended claims.
Although the present invention has been made detailed explanation and has quoted some specific embodiments as proof, to those skilled in the art, only otherwise leave that the spirit and scope of the present invention can be done various variations or correction is obvious.
Claims (7)
1. the cultural method of a microbial strain for degrading industrial waste water, this method may further comprise the steps:
A, aniline, phenol and nitrobenzene compounds in the trade effluent, preparation contains the liquid nutrient medium of the target compound of different concns gradient respectively, wherein aniline and nitrobenzene compounds are unique carbon source and nitrogenous source with it, phenol adds nitrogenous source in addition, also add needed other chemical matrix composition of microorganism strains growth in the described liquid nutrient medium, the partially liq substratum is changed into the selectivity solid medium;
B, the active sludge in the trade effluent joined carry out enrichment in the beef extract-peptone liquid nutrient medium, the liquid nutrient medium that is inoculated into the target compound that contains the different concns gradient is then tamed cultivation, culture temperature is 15-45 ℃, incubation time 6-25 days, domestication is cultivated 2-12 time repeatedly, the bacterial strain of domestication cultivation is coated the selectivity solid medium carry out the plate streaking separation, and carry out plasmid extraction, obtaining is sole carbon source and nitrogenous source with aniline or nitrobenzene compound respectively, with phenol be sole carbon source contain the plasmid bacterial strain;
C, the pseudomonas strain is inoculated into carries out enrichment in the beef extract-peptone liquid nutrient medium, be made into competent cell after the enrichment, the three kinds of plasmid bacterial strains and the described competent cell that obtain among the step B are resuspended in the LB liquid nutrient medium, are to hatch 10-45 hour under 10-40 ℃ the condition in temperature; The liquid nutrient medium that joins three kinds of target compounds that contain the different concns gradient is again tamed cultivation, the domestication culture temperature is 15-45 ℃, domestication incubation time 6-25 days, domestication is cultivated 2-12 time repeatedly, domestication is coated the selectivity solid medium after cultivating, be to hatch 10-45 hour under 10-40 ℃ the condition in temperature, carry out plasmid and degradation capability then and identify;
D, with after obtaining bacterial strain among the step C and carrying out purifying, promptly obtain to contain the microbial strain for degrading industrial waste water of plasmid.
2. the cultural method of microbial strain for degrading industrial waste water according to claim 1 is characterized in that: the nitrogenous source in the phenol liquid nutrient medium described in the steps A is that concentration is that 500mg-1500mg/L ammonium nitrate or concentration are a kind of in the 500mg-1500mg/L peptone.
3. the cultural method of microbial strain for degrading industrial waste water according to claim 1 and 2, it is characterized in that: other chemical matrix component is in the liquid nutrient medium described in the steps A: saltpetre: 0.5-1.5g/L; Sodium-chlor: 0.1-1g/L; K
2HPO
43H
2O:0.2g-0.8g/L; MgSO
47H
2O:0.2g-0.8g/L; FeSO
47H
2O0.005-0.015g/L.
4. the cultural method of microbial strain for degrading industrial waste water according to claim 1 is characterized in that: the culture temperature that domestication is cultivated among the step B is 20-40 ℃, and incubation time 7-21 days, domestication was cultivated 3-10 time repeatedly.
5. the cultural method of microbial strain for degrading industrial waste water according to claim 1 is characterized in that: among the step C in the LB liquid nutrient medium incubation temperature be 15-35 ℃, incubation time is 12-36 hour.
6. the cultural method of microbial strain for degrading industrial waste water according to claim 1 is characterized in that: the culture temperature that domestication is cultivated among the step C is 20-40 ℃, and incubation time 7-21 days, domestication was cultivated 3-10 time repeatedly.
7. according to the cultural method of claim 1 or 5 or 6 described microbial strain for degrading industrial waste water, it is characterized in that: to coat selectivity solid medium incubation temperature be 15-35 ℃ to bacterial strain among the step C, and incubation time is 12-36 hour.
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| CN110003990A (en) * | 2019-04-25 | 2019-07-12 | 南京巨鲨显示科技有限公司 | A kind of medical surface cleaning agent of microbiocidal and preparation method thereof |
| CN114437979A (en) * | 2022-02-14 | 2022-05-06 | 恒臻(无锡)生物科技有限公司 | Arthrobacter capable of degrading erucamide, and acquisition method, culture method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102515366A (en) * | 2011-12-23 | 2012-06-27 | 重庆文泰节能环保科技有限公司 | Microbiological degradation method for nitrobenzene industrial wastewater |
| CN102515366B (en) * | 2011-12-23 | 2013-07-10 | 重庆文泰节能环保科技有限公司 | Microbiological degradation method for nitrobenzene industrial wastewater |
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| CN102649943B (en) * | 2012-03-28 | 2013-03-13 | 哈尔滨师范大学 | Bacterium for high-efficiently degrading aniline under low-temperature condition |
| CN110003990A (en) * | 2019-04-25 | 2019-07-12 | 南京巨鲨显示科技有限公司 | A kind of medical surface cleaning agent of microbiocidal and preparation method thereof |
| CN114437979A (en) * | 2022-02-14 | 2022-05-06 | 恒臻(无锡)生物科技有限公司 | Arthrobacter capable of degrading erucamide, and acquisition method, culture method and application thereof |
| CN114437979B (en) * | 2022-02-14 | 2023-10-24 | 恒臻(无锡)生物科技有限公司 | Arthrobacter capable of degrading erucamide, and acquisition method, culture method and application thereof |
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| CN101701189B (en) | 2012-11-07 |
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